Research in Molecular Medicine http://rmm.mazums.ac.ir
Research in Molecular Medicine - Journal articles for year 2017, Volume 5, Number 2Yektaweb Collection - http://www.yektaweb.comen2017/5/11Viable but Non-culturable Bacteria: Clinical Practice and Future Perspectivehttp://rmm.mazums.ac.ir/browse.php?a_id=244&sid=1&slc_lang=en
In this editorial, the importance of VBNC in clinical practice and future researches interests has been pinpointed. Accurate detection of VBNC, especially for major mentioned pathogens, is the future research gap.&nbsp;Amin Talebi Bezmin AbadiEvaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCRhttp://rmm.mazums.ac.ir/browse.php?a_id=238&sid=1&slc_lang=en
<div style="text-align: justify;"><a name="OLE_LINK212"></a><strong>Background:</strong> The analysis of antigen-specific cytokine expression has been considered to evaluate the immune responses and vaccines efficacy in recent years. The aim of this study was to compare the cell-mediated immune response characteristics of two Capri pox virus (CaPV) vaccines against lumpy skin disease in cattle.<br>
<strong>Materials and Methods:</strong> Two Capri pox virus vaccines were administered to dairy cows of two farms and followed up to 5 weeks post vaccination. These vaccines were live attenuated Goat pox virus (GTP) Gorgan strain (n=20) and Sheep pox virus (SPP) Romanian strain (n=20). Cell-mediated immune response of vaccinated calves was evaluated using in vitro lymphocyte proliferation and IFN-g and IL-4 release assay after stimulation with recall vaccine strains, and in vivo cytokine expression in PBMCs by real-time PCR.<br>
<strong>Results:</strong> Lymphocyte proliferation in GTP- and SPP-vaccinated groups began to increase till reached to its peak at third week post vaccination and then decreased in the weeks thereafter. Stimulation index in stimulated PBMCs in GTP-vaccinated calves was higher than SPP-vaccinated calves in all weeks, which indicated higher levels of immunogenicity produced by the GTP-vaccine in cattle. Also, in both vaccinated groups the peak release of IFN-g and IL-4 proteins in cultured PBMCs in response to recall antigen was detected at week 3 post vaccination. Although the mean of the cytokine release in GTP-vaccinated calves was higher than SPP-vaccinated calves in all weeks of experiment, a significant difference was only observed at week 3 post vaccination (P<0.05). In contrast, the IFN-g mRNA expression in PBMCs of vaccinated groups was induced early, peaked at week 1 post vaccination and decreased in the weeks thereafter, and this rate was higher in GTP-vaccinated calves compared with SPP-vaccinated calves in all weeks, but the significant difference was only found at week 3 post vaccination (P<0.05). However, the IL-4 mRNA expression showed delayed induction and peaked at week 3, and unlike the SPP group, it remained at this level in GTP group, until the end of experiment. Also this rate of expression in GTP-vaccinated calves was higher than SPP-vaccinated calves in all weeks and had a significant increase at week 5 post vaccination (P<0.05).<br>
<strong>Conclusion:</strong> The findings show that due to induction of high level cell-mediated immune response in live attenuated GTP vaccine compared to SPP vaccine, GTP vaccine has a good immunogenic response, and therefore can be a better choice for vaccination against lumpy skin disease.<br>
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Nahideh Afzal ahangaranEffect of Lamium Album on Mitochondrial Oxidative Stress in Diabetic Ratshttp://rmm.mazums.ac.ir/browse.php?a_id=231&sid=1&slc_lang=en
<p style="text-align: justify;"><strong>Background:</strong> Diabetes mellitus (DM) is characterized by the presence of hyperglycemia. It has been documented that oxidative stress and reactive oxygen species (ROS) production have a key role in the pathogenesis of diabetes and its complications. Neutrophils as a part of immune system produce ROS, neutrophils function might be altered in diabetes. <em>Lamium album</em> is known to have antioxidant, and free radical scavenging actions. The aim of the present study was to evaluate the potential effect of <em>L. album</em> on mitochondrial ROS production from circulating neutrophils in diabetic rats.<br>
<strong>Materials and Methods: </strong>Twenty-one male Wistar rats were randomly divided into three groups: normal control rats receiving daily saline; diabetic control rats receiving daily saline; and diabetic rats treated daily with hydroalcoholic extract of <em>L. album </em>(100 mg/kg) for 28 days. On the 28<sup>th</sup>day of treatment, whole blood samples were obtained and mitochondrial ROS of neutrophils were measured by dihydrorhodamine (DHR) flow cytometric method. Also, fasting blood sugar (FBS) was measured.<br>
<strong>Results:</strong> Mitochondrial ROS didn&rsquo;t show any significant differences among diabetic rats treated with <em>L. album</em> extract, diabetic control rats, and normal control rats (P=0.8). Serum glucose in diabetic control was significantly higher than normal control rats (P=0.0001). However,<em> L. album</em> caused a remarkable decrease in serum glucose of diabetic rats (P=0.03).<br>
<strong>Conclusion:</strong> According to the present findings, it seems that <em>L. album</em> at a dose of 100 mg/kg could not decrease mitochondrial ROS production from neutrophils in diabetic rats. Further studies considering higher concentrations of <em>L. album</em> are appreciated to evaluate its impact on the production of mitochondrial ROS along with extracellular ROS in diabetes condition.</p>
Reza Jafari-ShakibChemical Composition and Antibacterial Effect of Medicinal Plants against Some Food-Borne Pathogenhttp://rmm.mazums.ac.ir/browse.php?a_id=242&sid=1&slc_lang=en
<div style="text-align: justify;"><strong>Background:</strong> <em>Pulicaria gnaphalodes</em>, <em>Ducrosia anethifolia</em>, <em>Trachyspermum copticum</em>, <em>Foeniculum vulgare</em> Mill and <em>Majorana hortensis</em> Minch are widely used as herbal plants in traditional medicine and they have been reported to have a variety of therapeutic effects. This study was carried out to evaluate the antimicrobial effects of essential oils (EOs) extracted from these medicinal herbs against six species of food-borne microorganisms.<br>
<strong>Materials and Methods:</strong> The EOs were analyzed by gas chromatography mass spectrometry (GC/MS). The detection of inhibitory effect of the EOs on the tested bacteria was carried out by agar disk-diffusion method and then MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) of the EOs against six bacteria were determined.<br>
<strong>Results:</strong> The analysis of the components of the essential oils (EOs) extracted by gas chromatography spectrometry allowed the identification of 63 compounds of the five tested EOs. All of these five tested EOs indicated an antimicrobial effect against strains of Bacillus sp and <em>Listeria Monocytogenes</em> ATCC1297. Essential oils from <em>T. copticum</em>, <em>M. hortensis</em> and <em>F. vulgare</em> possessed a wide spectrum of antibacterial activity against the growth of the six bacteria with zone diameter of inhibition (ZDI) between 14-32 mm, depending on the susceptibility of the tested organism.<br>
<strong>Conclusion: </strong>Antibacterial efficacy shown by these plants provides a scientific basis and thus validates their use as medicinal remedies. Isolation and purification of different phytochemicals may further yield significant antibacterial agents.</div>
Hassan HabibiIsolation and Identification of Phenanthrene-degrading Bacteria and Increasing the Biodegrading Ability by Synergistic Relationshiphttp://rmm.mazums.ac.ir/browse.php?a_id=212&sid=1&slc_lang=en
<div class="s13" style="margin-top: 0px; margin-right: 18px; margin-bottom: 0px; line-height: 1.8; text-align: justify; color: rgb(0, 0, 0); font-family: -webkit-standard; font-size: 18px;"><span class="s9" style="line-height: 14.399999618530273px; font-size: 12px; font-family: 'Times New Roman'; font-weight: bold;"><span class="bumpedFont15" style="line-height: 21.600000381469727px; font-size: 1.5em;">Background:</span></span><span class="s10" style="line-height: 14.399999618530273px; font-size: 12px; font-family: 'Times New Roman';"><span class="bumpedFont15" style="line-height: 21.600000381469727px; font-size: 1.5em;">&nbsp;</span></span>Polycyclic aromatic hydrocarbons are a large group of oil contaminants with carcinogenic, mutagenic and teratogenic effects. The release of these compounds in soil destroys animals, plants and microbial diversity and has several negative impacts on physical properties of the soil including the destruction of soil aggregates reduction in pores, and increase in soil bulk density. Many strains of microorganisms isolated have the phenanthrene-degrading ability but this study focused on isolation and identification of a phenanthrene-degrader bacterium for bioremediation of contaminated soils.</div>
<p class="s13" style="margin-top: 0px; margin-right: 18px; margin-bottom: 0px; line-height: 1.8; text-align: justify; color: rgb(0, 0, 0); font-family: -webkit-standard; font-size: 18px;"><strong>Background:</strong> Polycyclic aromatic hydrocarbons are a large group of oil contaminants with carcinogenic, mutagenic and teratogenic effects. The release of these compounds in soil destroys animals, plants and microbial diversity and has several negative impacts on physical properties of the soil including the destruction of soil aggregates reduction in pores, and increase in soil bulk density. Many strains of microorganisms isolated have the phenanthrene-degrading ability but this study focused on isolation and identification of a phenanthrene-degrader bacterium for bioremediation of contaminated soils.<br>
<strong>Materials and Methods:</strong> Enrichment technique was used for isolation and the most effective isolates, were named <em>pseudomonas aeruginosa</em> ZF1 and <em>Serratia marcescens</em> ZF2. The degradation experiments were conducted in the mineral salt medium (MSM) containing phenanthrene as the sole source of carbon and energy. The selection was based on phenanthrene biodegradation abilities. The isolates were identified using morphological, biochemical tests and 16S rDNA sequencing and after 10 days&rsquo; incubation at 30 &deg;C and pH = 7, the bacterial growth and Phe-degrading rate were evaluated by protein assay (Bradford) and gas chromatography (GC), respectively.<br>
<strong>Results:</strong> Biochemical tests and 16s rDNA gene sequence analysis revealed that isolated bacteria are similar to <em>Pseudomonas aeruginosa</em> ZF1 and <em>Serratia marcescens</em> ZF2 with 99% similarity. The results showed a mixture of ZF1 and ZF2 bacteria could degrade 83% at minimum concentrations of 200 ppm of phenanthrene whereas single strain culture of two bacteria had poor degradation abilities (less than 15%).<br>
<span class="s9" style="line-height: 14.399999618530273px; font-size: 12px; font-family: 'Times New Roman'; font-weight: bold;"><span class="bumpedFont15" style="line-height: 21.600000381469727px; font-size: 1.5em;">Conclusion</span></span><span class="s10" style="line-height: 14.399999618530273px; font-size: 12px; font-family: 'Times New Roman';"><span class="bumpedFont15" style="line-height: 21.600000381469727px; font-size: 1.5em;">:&nbsp;</span></span>Results showed that isolated co-culture bacteria have high potential to degrade phenanthrene with the best results achieved when the enriched consortium was used and this mixture was shown to be an appropriate candidate for bioremediation purposes.</p>
Zahra FathiBioinformatics Designing of 10-23 Deoxyribozyme against Coding Region of Beta-galactosidase Gene http://rmm.mazums.ac.ir/browse.php?a_id=240&sid=1&slc_lang=en
<div style="text-align: justify;"><strong>Background: </strong>Deoxyribozymes (Dzs) can play a role as gene expression inhibitors at mRNA level. Among Dzs, the 10-23 deoxyribozyme has significant potentials for treatment of diseases. Designed Dz includes a catalytic core made of 15 deoxyribonucleotides and two binding arms consisted of 6-12 nucleotides for site specific binding to target RNA and hydrolysis. The enzyme has characteristic features for cleavage of the RNA target between an unpaired purine (A, G) and a paired pyrimidine (C or U). In this study, 10-23 Dz is designed for the coding region of the &alpha;-peptide of a <em>lacZ</em> gene.</div>
<div style="text-align: justify;"><strong>Material and Methods: </strong>The primary sequence of a plasmid with &alpha;-complementation ability was taken from addgene database. To confirm sequence validity, ExPASy was used to analyze related ORFs for the retrieved sequence. The ORF with identical sequence to &alpha;-peptide was selected in the reverse complement sequence. Subsequently, the secondary structure of the &alpha;-peptide was analyzed in DINAMelt web server and Mfold software. Then the intended target site was selected inside the coding region of the &alpha;-peptide. The Dzs sequence was designed for the target site with nucleotide binding arms.<br>
<strong>Results and conclusion:</strong> The resulted Dz in this study can be used as a promising catalytic DNA inside bacterial cells for blue-white screening. Criteria such as biological stability and catalytic rate of such enzymes must be evaluated in vivo and in vitro.</div>
Abolghasem EsmaeiliIdentification of a Neonate with Thalassemia Intermedia Despite Premarital Screening Program in Mazandaran Province (Co-inheritance of Hb Knossos and IVS II-1 G> A Mutations)http://rmm.mazums.ac.ir/browse.php?a_id=228&sid=1&slc_lang=en
<p style="text-align: justify;"><strong>Background:</strong> Beta thalassemia is a common health problem in Iran especially in Northern provinces. Premarital screening for thalassemia is compulsory in Iran and identification of the carriers is based on primary CBC (Cell Blood Count) and hemoglobin electrophoresis. Silent mutations on &beta;-globin gene have borderline or normal hematological indices that cannot be detected in premarital screening.<br>
<strong>Material and Methods:</strong> A 4 years old boy affected with &beta;-thalassemia was referred to the lab for molecular analysis. During the screening program for &beta;-thalassemia, his father and mother were diagnosed as &alpha; and &beta;-thalassemia carrier respectively.&nbsp;<br>
<strong>Results:</strong> The results of molecular analysis showed that in addition to the &alpha;-globin single gene deletion (&alpha;<sup>3.7</sup>) the father have also carried a silent mutation on his &beta;-globin gene named HBB c.82G>T or Hb <sup>Knossos</sup> that was missed in screening program.<br>
<strong>Conclusion:</strong> The presented case shows that using CBC and hemoglobin electrophoresis in premarital screening program for detecting &beta;-thalassemia carriers is not a valid approach and individuals who are carriers of silent &beta;-globin gene mutations are missed in this procedure. Hence, in premarital screening program precise molecular investigation especially when the partner is a typical &beta;-thalassemia carrier is recommended.</p>
Mohammad Reza Mahdavi